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Live cell analyses of synaptonemal complex dynamics and chromosome movements in cultured mouse testis tubules and embryonic ovaries

During mammalian meiotic prophase, homologous chromosomes connect through the formation of the synaptonemal complex (SC). SYCP3 is a component of the lateral elements of the SC. We have generated transgenic mice expressing N- or C-terminal fluorescent-tagged SYCP3 (mCherry-SYCP3 (CSYCP) and SYCP3-mC...

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Autores principales: Enguita-Marruedo, Andrea, Van Cappellen, Wiggert A., Hoogerbrugge, Jos W., Carofiglio, Fabrizia, Wassenaar, Evelyne, Slotman, Johan A., Houtsmuller, Adriaan, Baarends, Willy M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6096571/
https://www.ncbi.nlm.nih.gov/pubmed/29582139
http://dx.doi.org/10.1007/s00412-018-0668-7
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author Enguita-Marruedo, Andrea
Van Cappellen, Wiggert A.
Hoogerbrugge, Jos W.
Carofiglio, Fabrizia
Wassenaar, Evelyne
Slotman, Johan A.
Houtsmuller, Adriaan
Baarends, Willy M.
author_facet Enguita-Marruedo, Andrea
Van Cappellen, Wiggert A.
Hoogerbrugge, Jos W.
Carofiglio, Fabrizia
Wassenaar, Evelyne
Slotman, Johan A.
Houtsmuller, Adriaan
Baarends, Willy M.
author_sort Enguita-Marruedo, Andrea
collection PubMed
description During mammalian meiotic prophase, homologous chromosomes connect through the formation of the synaptonemal complex (SC). SYCP3 is a component of the lateral elements of the SC. We have generated transgenic mice expressing N- or C-terminal fluorescent-tagged SYCP3 (mCherry-SYCP3 (CSYCP) and SYCP3-mCherry (SYCPC)) to study SC dynamics and chromosome movements in vivo. Neither transgene rescued meiotic aberrations in Sycp3 knockouts, but CSYCP could form short axial element-like structures in the absence of endogenous SYCP3. On the wild-type background, both fusion proteins localized to the axes of the SC together with endogenous SYCP3, albeit with delayed initiation (from pachytene) in spermatocytes. Around 40% of CSYCP and SYCPC that accumulated on the SC was rapidly exchanging with other tagged proteins, as analyzed by fluorescent recovery after photobleaching (FRAP) assay. We used the CSYCP transgenic mice for further live cell analyses and observed synchronized bouquet configurations in living cysts of two or three zygotene oocyte nuclei expressing CSYCP, which presented cycles of telomere clustering and dissolution. Rapid chromosome movements were observed in both zygotene oocytes and pachytene spermatocytes, but rotational movements of the nucleus were more clear in oocytes. In diplotene spermatocytes, desynapsis was found to proceed in a discontinuous manner, whereby even brief chromosome re-association events were observed. Thus, this live imaging approach can be used to follow changes in the dynamic behavior of the nucleus and chromatin, in normal mice and different infertile mouse models. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00412-018-0668-7) contains supplementary material, which is available to authorized users.
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spelling pubmed-60965712018-08-24 Live cell analyses of synaptonemal complex dynamics and chromosome movements in cultured mouse testis tubules and embryonic ovaries Enguita-Marruedo, Andrea Van Cappellen, Wiggert A. Hoogerbrugge, Jos W. Carofiglio, Fabrizia Wassenaar, Evelyne Slotman, Johan A. Houtsmuller, Adriaan Baarends, Willy M. Chromosoma Original Article During mammalian meiotic prophase, homologous chromosomes connect through the formation of the synaptonemal complex (SC). SYCP3 is a component of the lateral elements of the SC. We have generated transgenic mice expressing N- or C-terminal fluorescent-tagged SYCP3 (mCherry-SYCP3 (CSYCP) and SYCP3-mCherry (SYCPC)) to study SC dynamics and chromosome movements in vivo. Neither transgene rescued meiotic aberrations in Sycp3 knockouts, but CSYCP could form short axial element-like structures in the absence of endogenous SYCP3. On the wild-type background, both fusion proteins localized to the axes of the SC together with endogenous SYCP3, albeit with delayed initiation (from pachytene) in spermatocytes. Around 40% of CSYCP and SYCPC that accumulated on the SC was rapidly exchanging with other tagged proteins, as analyzed by fluorescent recovery after photobleaching (FRAP) assay. We used the CSYCP transgenic mice for further live cell analyses and observed synchronized bouquet configurations in living cysts of two or three zygotene oocyte nuclei expressing CSYCP, which presented cycles of telomere clustering and dissolution. Rapid chromosome movements were observed in both zygotene oocytes and pachytene spermatocytes, but rotational movements of the nucleus were more clear in oocytes. In diplotene spermatocytes, desynapsis was found to proceed in a discontinuous manner, whereby even brief chromosome re-association events were observed. Thus, this live imaging approach can be used to follow changes in the dynamic behavior of the nucleus and chromatin, in normal mice and different infertile mouse models. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00412-018-0668-7) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2018-03-26 2018 /pmc/articles/PMC6096571/ /pubmed/29582139 http://dx.doi.org/10.1007/s00412-018-0668-7 Text en © The Author(s) 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Original Article
Enguita-Marruedo, Andrea
Van Cappellen, Wiggert A.
Hoogerbrugge, Jos W.
Carofiglio, Fabrizia
Wassenaar, Evelyne
Slotman, Johan A.
Houtsmuller, Adriaan
Baarends, Willy M.
Live cell analyses of synaptonemal complex dynamics and chromosome movements in cultured mouse testis tubules and embryonic ovaries
title Live cell analyses of synaptonemal complex dynamics and chromosome movements in cultured mouse testis tubules and embryonic ovaries
title_full Live cell analyses of synaptonemal complex dynamics and chromosome movements in cultured mouse testis tubules and embryonic ovaries
title_fullStr Live cell analyses of synaptonemal complex dynamics and chromosome movements in cultured mouse testis tubules and embryonic ovaries
title_full_unstemmed Live cell analyses of synaptonemal complex dynamics and chromosome movements in cultured mouse testis tubules and embryonic ovaries
title_short Live cell analyses of synaptonemal complex dynamics and chromosome movements in cultured mouse testis tubules and embryonic ovaries
title_sort live cell analyses of synaptonemal complex dynamics and chromosome movements in cultured mouse testis tubules and embryonic ovaries
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6096571/
https://www.ncbi.nlm.nih.gov/pubmed/29582139
http://dx.doi.org/10.1007/s00412-018-0668-7
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